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Bulletin of Botanical Research ›› 2005, Vol. 25 ›› Issue (4): 436-440.doi: 10.7525/j.issn.1673-5102.2005.04.013

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Cloning of human granulocyte-macrophage colony stimulating factor (hGM-CSF) gene in Anabaena sp. PCC7120

WEI Lan-Zhen1, Jin Rui1, MA Wei-Min2, SHI Ding-Ji1,3, Gan Ren-Bao4, WANG Quan-Xi1   

  1. 1. Department of Biology, Shanghai Normal University, Shanghai 200234;
    2. Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Graduate School of the Chinese Academy of Sciences, Shanghai 200032;
    3. Institute of Botany, Chinese Academy of Science, Beijing 100093;
    4. Institute of Biochemistry, Chinese Academy of Science, Shanghai 200031
  • Received:2004-11-24 Online:2005-12-15 Published:2016-06-14

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Abstract: Human granulocyte-macrophage colony stimulating factor(hGM-CSF) is one of a family glycoprotein cytokines that have potent effects in stimulating proliferation, maturation and function of hematopoietic cells.The gene encoding hGM-CSF was PCR amplified from the plasmid pAGMT-8 and modified by addition SD sequence to promote prokaryotic expression.The resultant hGM-CSF was inserted into downstream of the strong promoter, PpsbA of expression vector pRL-439, then ligated with pDC-08 to produce the shuttle expression vector, pDC-GM.The resulting pDC-GM was transferred into the filamentous, heterocystour cyanobacterium, Anabaena PCC7120, by the tri-parental conjugation transfer method.When cells were cultivated without antibiotics for many generations, the transgenic cyanobacteria remained resistant to neomycin.PCR amplification of wild type and transgenic Anabaena cells with primers P1 and P2 revealed no band in wild type and plasmid free cells, whereas transgenic cells yielded a about(390 bp) band.This is the first report of expression hGM-CSF in cyanobacteria cells.

Key words: Human granulocyte-macrophage colony stimulating factor, triparental conjugative transfer, transgenic cyanobacterium, Anabaena sp.PCC7120